Abstract
To evaluate the prospects for transposon mutagenesis in the autogamous diploid legume Lotus japonicus, the behaviour of the maize transposable element Ac was analysed in the progeny of 38 independent transgenic plants. The conditions for monitoring donor site excision using histochemical localization of β-glucuronidase activity or the alternative spectinomycin resistance assay were established, and used to follow Ac mobility through two generations. Somatic excision was monitored as variegated cotyledons in the T2 generation and germinal excision events were scored in segregating T3 families as complete β-glucuronidase-mediated staining of cotyledons or as a fully green spectinomycin-resistant phenotype. Using these assays an average germinal excision frequency of 12% was estimated in the T3 offspring from variegated plants. The fidelity of the excision assays was ascertained by comparing the frequency of germinal excision to the frequency of Ac reinsertion at new positions of the genome. Transposition of Ac in 42% of the plants and detection of the characteristic Ac insertion/excision footprints suggests that insertion mutagenesis with the autonomous maize Activator element is feasible in Lotus japonicus. Parameters influencing Ac behaviour, such as dosage, position effects and modification of the element itself, were also investigated comparing homozygous and hemizygous plants from the same family and by analysing different transformants.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- W:
-
white
- V:
-
variegated
- FG:
-
fully green
- FB:
-
fully blue
- aadA :
-
spectinomycin adenyltransferase
References
Aarts MGM, Dirkse WG, Stiekema WJ, Pereira A: Transposon tagging of a male sterility gene in Arabidopsis. Nature 363: 715–717 (1993).
Baker B, Coupland G, Fedoroff N, Starlinger P, Schell J: Phenotypic assay for excision of the maize controlling element Ac in tobacco. EMBO J 6: 1547–1554 (1987).
Bancroft I, Bhatt AM, Sjodin C, Scofield S, Jones JDG, Dean C: Development of an efficient two-element transposon taggmg system in Arabidopsis thaliana. Mol Gen Genet 233: 449–461 (1992).
Bancroft I, Jones JDG, Dean C: Heterologous transposon tagging of the DRL1 locus in Arabidopsis. Plant Cell 5: 631–638 (1993).
Caetano-Anollés G, Gresshoff PM: Plant genetic control of nodulation. Annu Rev Microbiol 45: 345–382 (1991).
Cardon GH, Frey M, Saedler H, Gierl A: Mobility of the maize transposable element En/Spm in Arabidopsis thaliana. Plant J 3: 773–784 (1993).
Chuck G, Robbins T, Nijjar C, Ralston E, Courtney-Gutterson N, Dooner HK: Tagging and cloning of a petunia flower color gene with the maize transposable element Activator. Plant Cell 5: 371–378 (1993).
Coen ES, Meyerowitz EM: The war of the whorls: genetic interactions controlling flower development. Nature 353: 31–37 (1991).
Coupland G, Baker B, Schell J, Starlinger P: Characterization of the maize transposable element Ac by internal deletions. EMBO J 7: 3653–3659 (1988).
Coupland G, Plum C, Chatterjee S, Post A, Starlinger P: Sequences near the termini are required for transposition of the maize transposon Ac in transgenic tobacco plants. Proc Natl Acad Sci USA 86: 9385–9388 (1989).
Dean C, Sjodin C, Bancroft I, Lawson E, Lister C, Scofield S, Jones J: Development of an efficient transposon tagging system in Arabidopsis thaliana. Symposia of the Society for Experimental Biology XLV: 63–75 (1991).
Dean C, Sjodin C, Page T, Jones J, Lister C: Behaviour of the maize transposable element Ac in Arabidopsis thaliana. Plant J 2: 69–81 (1992).
Döring H-P, Starlinger P: Molecular genetics of transposable elements in plants. Annu Rev Genet 20: 175–200 (1986).
Earp DJ, Lowe B, Baker B: Amplification of genomic sequences flanking transposable elements in host and heterologous plants: a tool for transposon tagging and genome characterization. Nucl Acids Res 18: 3271–3279 (1990).
Ellis JG, Finnegan EJ, Lawrence GJ: Developing a transposon tagging system to isolate rust-resistance genes from flax. Theor Appl Genet 85: 46–54 (1992).
English JJ, Harrison KA, Jones JDG: Agenetic analysis of DNA sequence requirements for Dissociation state I activity in tobacco. Plant Cell 5: 501–514 (1993).
Fedoroff NV, Wessler S, Shure M: Isolation of the transposable maize controlling elements Ac and Ds. Cell 35: 243–251 (1983).
Fedoroff NV: Maize transposable elements. In: Howe M, Berg D (eds) Mobile DNA, pp. 375–411. ASM Press, Washington, DC (1989).
Fedoroff NV, Smith DL: A versatile system for detecting transposition in Arabidopsis. Plant J 3: 273–289 (1993).
Franssen HJ, Vijn I, Yang WC, Bisseling T: Developmental aspects of the Rhizobium-legume symbiosis. Plant Mol Biol 19: 89–107 (1992).
Handberg K, Stougaard J: Lotus japonicus, an autogamous, diploid legume species for classical and molecular genetics. Plant J 2: 487–496 (1992).
Handberg K, Stiller J, Thykjaer T, Stougaard J: Transgenic plants: Agrobacterium mediated transformation of the diploid legume Lotus japonicus. In: Celis JE (ed) Cell Biology: A Laboratory Handbook, Academic Press, New York (1994).
Jefferson RA: Assaying chimeric genes in plants: the GUS fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Jones JDG, Carland FM, Maliga P, Dooner HK: Visual detection of transposition of the maize element Activator (Ac) in tobacco seedlings. Science 244: 204–207 (1989).
Jones JDG, Carland F, Lim E, Ralston E, Dooner HK: Preferential transposition of the maize element Activator to linked chromosomal locations in tobacco. Plant Cell 2: 701–707 (1990).
Jones JDG, Harper L, Carland F, Ralston E, Dooner HK: Reversion and altered variegation of an SPT:Ac allele in tobacco. Maydica 36: 329–335 (1991).
Jones JDG, Shlumukov L, Carland F, English J, Scofield SR, Bishop GJ, Harrison K: Effective vectors for transformation, expression of heterologous genes, and assaying transposition excision in transgenic plants. Transgenic Res 1: 285–297 (1992).
Kunze R, Starlinger P: The putative transposase of transposable element Ac from Zea mays L. interacts with subterminal sequences of Ac. EMBO J 8: 3177–3185 (1989).
Pohlmann RF, Fedoroff NV, Messing J: The nucleotide sequence of the maize controlling element Activator. Cell 37: 635–643 (1984).
Saedler H, Nevers P: Transposition in plants: a molecular model. EMBO J 4: 585–590 (1985).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Scofield SR, English JJ, Jones JGD: High level expression of the Activator transposase gene inhibits the excision of Dissociation in tobacco cotyledons. Plant Cell 5: 507–517 (1993).
Scofield SR, Jones DA, Harrison K, Jones JDG: Chloroplast targeting of spectinomycin adenyltransferase provides a cell autonomous marker for monitoring transposon excision in tomato and tobacco. Mol Gen Genet 244: 189–196 (1994).
Stougaard J: Substrate-dependent negative selection in plants using a bacterial cytosine deaminase gene. Plant J 3: 755–761 (1993).
VanSluys MA, Tempé J, Fedoroff N: Studies on the introduction and mobility of the maize Activator element in Arabidopsis thaliana and Daucus carota. EMBO J 6: 3881–3889 (1987).
Yang C-H, Ellis JG, Michelmore RW: Infrequent transposition of Ac in lettuce, Lectuca sativa. Plant Mol Biol 22: 793–805 (1993).
Yoder JI, Palys J, Alpert K, Lassner M: Ac transposition in transgenic tomato plants. Mol Gen Genet 213: 291–296 (1988).
Zhou J H, Atherly AG: In situ detection of transposition of the maize controlling element (Ac) in transgenic soybean tissues. Plant Cell Rep 8: 542–545 (1990).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Thykjaer, T., Stiller, J., Handberg, K. et al. The maize transposable element Ac is mobile in the legume Lotus japonicus . Plant Mol Biol 27, 981–993 (1995). https://doi.org/10.1007/BF00037025
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00037025